GSM7179082: Runx1+ IAHC; Mus musculus; RNA-Seq
E11 embryos were obtained from Runx1-GFP/+ males and females time pregnancy and AGM cells stained with NG2 Cy3 (1:100, Millipore), PDGFRβ APC (1:100, Biolegend), CD31 PECy7 (1:4000, eBioscience), CD45 PerCypCy5.5 (1:400, BD Pharmingen), ckit BV421 (1:500, BD Horizon) and CD45 PerCpCy5.5 (1:400, BD Pharmingen) for 30 min at 4oC, washed, centrifuged, then resuspended in PBS/FCS/PS for analysis and cell sorting. Cells were purified using BD FACS ARIA Fusion cell sorter (BD Bioscience) and Sytox AAD was used to select viable cells. Sorted cells were sorted and collected directly into 20μl of lysis buffer containing Nuclease-free water (Ambion AM9930) 0.2% Triton and 1/20 RNAse inhibitor (Thermoscientific 10777019). RNA libraries for RNA-seq were prepared using SMARTER mRNA-Seq Library Prep Kit following manufacturer's protocols.